Summary
The production of doubled haploids (DH) has become a necessary tool in advanced plant breeding institutes and commercial companies for breeding many crop species. However, the development of new, more efficient and cheaper large scale production protocols has meant that doubled haploids are also recently being applied in less advanced breeding programmes. This Manual was prepared to stimulate the wider use of this technology for speeding and opening up new breeding possibilities for may crops including some woody species. Since the construction of genetic maps using molecular markers requires the development of segregating doubled haploid populations in numerous crop species, we hope that this Manual will also help molecular biologists in establishing such mapping populations.
Requests from researchers and trainees for systematised protocols dealing with the production of doubled haploids in various species were instrumental in initiating work on this Manual, and the idea to prepare it was strongly supported by European scientists working under the EU large scale program 'COST 851' on 'Gametic cells and molecular breeding for crop improvement'. Many of the authors who provided protocols for the Manual participated in these activities, thereby helping to transfer DH technology in countries and over continents.
This is the first manual presenting a set of protocols for production of (DH) in 22 major crop plant species including 4 tree species. It contains various protocols and approaches of DH production proved for different germplasm of the same species. The protocols describe in detail all steps of DH production- from donor plant growth conditions, through in vitro procedures, media composition and preparation, to regeneration of haploid plants and chromosome doubling methods.The users of the Manual will be able to choose among microspore versus anther culture, wide hybridization or gynogenesis, the most suitable method for production of DH in particular laboratory conditions for their plant materials. The user will also find information on organisation of a doubled haploid laboratory, basic DH media and on associated simple cytogenetic methods for ploidy level analysis. The ractical protocols are supplemented with the list of published DH protocols for other crop plants and separate chapters dealing with major applications of DH in breeding, mutant production, transgenesis, and genetic mapping and genomics.